Long-term forskolin stimulation induces AMPK activation and thereby enhances tight junction formation in human placental trophoblast BeWo cells.
BeWo cells, derived from human choriocarcinoma, have been known to respond to forskolin or cAMP analogues by differentiating into multinucleated cells- like syncytiotrophoblasts on the surfaces of chorionic villi of the human placenta. In this study, we demonstrated that long-term treatment with forskolin enhances the tight junction (TJ) formation in human placental BeWo cells. Interestingly, AMPK activation and phosphorylation of acetyl-CoA carboxylase (ACC), a molecule downstream from AMPK, were induced by long-term incubation (>12h) with forskolin, despite not being induced by acute stimulation with forskolin. In addition, co-incubation with an AMPK inhibitor, compound C, as well as overexpression of an AMPK dominant negative mutant inhibited forskolin-induced TJ formation. Thus, although the molecular mechanism underlying AMPK activation via the forskolin stimulation is unclear, the TJ formation induced by forskolin is likely to be mediated by the AMPK pathway. Taking into consideration that TJs are present in the normal human placenta, this mechanism may be important for forming the placental barrier system between the fetal and maternal circulations.
Egawa
M
M
Department of Medical Chemistry, Division of Molecular Medical Science, Graduate School of Biomedical Science, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8551, Japan.
Kamata
H
H
Kushiyama
A
A
Sakoda
H
H
Fujishiro
M
M
Horike
N
N
Yoneda
M
M
Nakatsu
Y
Y
Ying
Guo
G
Jun
Zhang
Z
Tsuchiya
Y
Y
Takata
K
K
Kurihara
H
H
Asano
T
T
D055372:AMP-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D002822:Choriocarcinoma; D005576:Colforsin / Q000494:pharmacology*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D006801:Humans; D019426:Integrases / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D021041:Placental Circulation / Q000502:physiology; D011247:Pregnancy; D019108:Tight Junctions / Q000187:drug effects* / Q000378:metabolism*; D014162:Transfection; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D014594:Uterine Neoplasms
D005576:Colforsin; D008156:Luciferases; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; C045073:Cre recombinase; D019426:Integrases